Integrating multi-unit electrophysiology and plastic culture dishes for network neuroscience.
The electrophysiological characterisation of cultured neurons is of paramount importance for drug discovery, safety pharmacology and basic research in the neurosciences. Technologies offering low cost, low technical complexity and potential for scalability towards high-throughput electrophysiology on in vitro neurons would be advantageous, in particular for screening purposes. Here we describe a plastic culture substrate supporting low-complexity multi-unit loose-patch recording and stimulation of developing networks while retaining manufacturability compatible with low-cost and large-scale production. Our hybrid polydimethylsilane (PDMS)-on-polystyrene structures include chambers (6 mm in diameter) and microchannels (25 microm x 3.7 microm x 1 mm) serving as substrate-embedded recording pipettes. Somas are plated and retained in the chambers due to geometrical constraints and their processes grow along the microchannels, effectively establishing a loose-patch configuration without human intervention. We demonstrate that off-the-shelf voltage-clamp, current-clamp and extracellular amplifiers can be used to record and stimulate multi-unit activity with the aid of our dishes. Spikes up to 50 pA in voltage-clamp and 300 microV in current-clamp modes are recorded in sparse and bursting activity patterns characteristic of 1 week-old hippocampal cultures. Moreover, spike sorting employing principal component analysis (PCA) confirms that single microchannels support the recording of multiple neurons. Overall, this work suggests a strategy to endow conventional culture plasticware with added functionality to enable cost-efficient network electrophysiology.
Morales
Ricardo
R
Catalonia Bioengineering Institute, 08028, Barcelona, Spain.
Riss
Michael
M
Wang
Ling
L
Gavín
Rosalina
R
Del Río
Jose Antonio
JA
Alcubilla
Ramon
R
Claverol-Tinturé
Enric
E
D002245:Carbon Dioxide / Q000737:chemistry; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D003365:Costs and Cost Analysis; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004553:Electric Conductivity; D004594:Electrophysiology / Q000379:methods*; D005110:Extracellular Space / Q000378:metabolism; D008839:Microelectrodes; D044085:Microfluidics; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D009488:Neurosciences / Q000379:methods*; D018408:Patch-Clamp Techniques; D010969:Plastics / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D014162:Transfection
D004129:Dimethylpolysiloxanes; D010969:Plastics; D011137:Polystyrenes; D002245:Carbon Dioxide; C013830:baysilon